Cloning and characterization of a recombinant Rmi-serpin expressed in Pichia pastoris yeast

Rodriguez-Valle, Manuel, Xu, Tao, Vance, Megan and Lew-Tabor, Ala (2011). Cloning and characterization of a recombinant Rmi-serpin expressed in Pichia pastoris yeast. In: TTP7: 7th Tick and Tick-Borne Pathogen International Conference. Proceedings. Tick and Tick-borne Pathogen International Conference (7th, TTP, 2011), Zaragoza, Spain, (65-65). 28 August-2 September 2011.

Author Rodriguez-Valle, Manuel
Xu, Tao
Vance, Megan
Lew-Tabor, Ala
Title of paper Cloning and characterization of a recombinant Rmi-serpin expressed in Pichia pastoris yeast
Formatted title
Cloning and characterization of a recombinant Rmi-serpin expressed in Pichia pastoris yeast
Conference name Tick and Tick-borne Pathogen International Conference (7th, TTP, 2011)
Conference location Zaragoza, Spain
Conference dates 28 August-2 September 2011
Proceedings title TTP7: 7th Tick and Tick-Borne Pathogen International Conference. Proceedings
Publication Year 2011
Sub-type Published abstract
Start page 65
End page 65
Total pages 1
Collection year 2012
Formatted Abstract/Summary
Rhipicephalus (Boophilus) microplus is a very important bovine ectoparasite widely distributed in tropical and subtropical regions of the world (Lew-Tabor et al 2009). Its success as an ectoparasite is associated with its capacity to disarm the antihemostatic and antiinflammatory reactions of host using protease inhibitors among other proteins (Rodriguez-Valle et al 2010).The saliva of Ixodid ticks have anticoagulant, antihemostatic, antiinflammatory, immunosuppressive and anticomplement activities. Serpins are a protease inhibitor family with an important role in the modulation of host – parasite interactions. We describe here the expression of a recombinant Rmi Serpin in Pichia pastoris (GSS115 strain). The Rmi-serpin gene was cloned into pPICalpha-A vector and was secreted into the culture media after methanol induction at 5 mg L-1. The recombinant Rmi-serpin was classified using bioinformatics as a homologue of clade B serine inhibitors (Hit ID: NP_001099089, 33%). The expression of this Rmi-serpin in the salivary gland was corroborated by qRT-PCR. Trypsin inhibition assay was carried out for determining the capacity of Rmi-serpin to inhibit serine protease. Recombinant Rmi-serpin from P. pastoris highly inhibited the trypsin activity with a molar ration of 1:1 (protein-protein). Further functional studies of Rmi-serpins will be conducted.
Keyword Serpin
Protease inhibitor
Tick
R. microplus
Q-Index Code EX
Q-Index Status Provisional Code
Institutional Status UQ

Document type: Conference Paper
Collections: Queensland Alliance for Agriculture and Food Innovation
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Created: Wed, 19 Oct 2011, 15:54:08 EST by Dr Alicja Lew-tabor on behalf of Qld Alliance for Agriculture and Food Innovation